Network Pharmacology and Experimental Verification Strategies to Illustrate the Mechanism of Jian-Pi-Yi-Shen Formula in Suppressing Epithelial–Mesenchymal Transition

Abstract

Jian-Pi-Yi-Shen formula (JPYSF), a traditional Chinese medicine, has been recommended to treat renal fibrosis for decades. Previous studies had shown that JPYSF could inhibit epithelial–mesenchymal transition (EMT), an important regulatory role in renal fibrosis. However, the mechanism of JPYSF action is largely unknown. In this study, network pharmacology and experimental verification were combined to elucidate and identify the potential mechanism of JPYSF against renal fibrosis by suppressing EMT at molecular and pathway levels. Network pharmacology was first performed to explore the mechanism of JPYSF against renal fibrosis targeting EMT, and then a 5/6 nephrectomy (5/6 Nx)-induced rat model of renal fibrosis was selected to verify the predictive results by Masson’s trichrome stains and western blot analysis. Two hundred and thirty-two compounds in JPYSF were selected for the network approach analysis, which identified 137 candidate targets of JPYSF and 4,796 known therapeutic targets of EMT. The results of the Gene Ontology (GO) function enrichment analysis included 2098, 88, and 133 GO terms for biological processes (BPs), molecular functions (MFs), and cell component entries, respectively. The top 10 enrichment items of BP annotations included a response to a steroid hormone, a metal ion, oxygen levels, and so on. Cellular composition (CC) is mainly enriched in membrane raft, membrane microdomain, membrane region, etc. The MF of JPYSF analysis on EMT was predominately involved in proximal promoter sequence-specific DNA binding, protein heterodimerization activity, RNA polymerase II proximal promoter sequence-specific DNA binding, and so on. The involvement signaling pathway of JPYSF in the treatment of renal fibrosis targeting EMT was associated with anti-fibrosis, anti-inflammation, podocyte protection, and metabolism regulation. Furthermore, the in vivo experiments confirmed that JPYSF effectively ameliorated interstitial fibrosis and inhibited the overexpression of α-SMA, Wnt3a, and β-catenin, and increased the expression of E-cadherin by wnt3a/β-catenin signaling pathway in 5/6 Nx-induced renal fibrosis rats. Using an integrative network pharmacology-based approach and experimental verification, the study showed that JPYSF had therapeutic effects on EMT by regulating multi-pathway, among which one proven pathway was the Wnt3a/β-catenin signaling pathway. These findings provide insights into the renoprotective effects of JPYSF against EMT, which could suggest directions for further research of JPYSF in attenuating renal fibrosis by suppressing EMT.

Jian-Pi-Yi-Shen formula alleviates renal fibrosis by restoring NAD+ biosynthesis in vivo and in vitro

“…Studies have shown that JPYSF protected kidney from fibrotic injury [12,13,51]. Some active ingredients in JPYSF have been reported to exert anti-fibrotic effects through acting on cellular signal pathways, such as inhibiting the TGF-β1 pathway, activating Sirt1-mediated autophagy, and inhibiting the Wnt3a/β-catenin pathway [52][53][54][55][56]. Our data suggested that JPYSF alleviates renal fibrosis by restoring NAD + biosynthesis.…”

Heidihuangwan alleviates renal fibrosis in rats with 5/6 nephrectomy by inhibiting autophagy

“…It has been shown that single herbs (Liu et al, 2022;Qian et al, 2022), herbal extracts (Yu, 2020;Zheng et al, 2021;Cong et al, 2022;Ma et al, 2022), and Chinese medicine compounds (Chen et al, 2022a;Chen et al, 2022b;Wu et al, 2022;Zhao et al, 2022) have made great progress in the prevention and even treatment of renal fibrosis (Xu et al, 2021). They regulate multiple signaling pathways such as Wnt/β-catenin (Zhao et al, 2022), TGF-β1/Smad (Mao et al, 2019;Wu et al, 2022), Akt/mTOR (Lian et al, 2022), CX3CL1-RAF/MEK/ ERK (Hu et al, 2022b), mTORC1/p70S6K (Chen et al, 2019), SIRT1-NF-κB (Wang et al, 2018b), and interfere with different cytokine or inflammatory factors expression to regulate the development and progression of renal fibrosis.…”

“…It has been shown that single herbs (Liu et al, 2022;Qian et al, 2022), herbal extracts (Yu, 2020;Zheng et al, 2021;Cong et al, 2022;Ma et al, 2022), and Chinese medicine compounds (Chen et al, 2022a;Chen et al, 2022b;Wu et al, 2022;Zhao et al, 2022) have made great progress in the prevention and even treatment of renal fibrosis (Xu et al, 2021). They regulate multiple signaling pathways such as Wnt/β-catenin (Zhao et al, 2022), TGF-β1/Smad (Mao et al, 2019;Wu et al, 2022), Akt/mTOR (Lian et al, 2022), CX3CL1-RAF/MEK/ ERK (Hu et al, 2022b), mTORC1/p70S6K (Chen et al, 2019), SIRT1-NF-κB (Wang et al, 2018b), and interfere with different cytokine or inflammatory factors expression to regulate the development and progression of renal fibrosis. The Chinese herbal extract AS-IV, which has been widely recognized to have significant anti-nephrogenic effects (Yu, 2020;Fu et al, 2022), is the main active component of Huangqi, a Chinese herbal medicine, and its mechanism has been richly studied and Frontiers in Pharmacology frontiersin.org repeatedly validated (Wang et al, 2018b;Chen et al, 2019;Mao et al, 2019;Hu et al, 2022b;Lian et al, 2022).…”

The potential mechanism of huazhuojiedu decoction in the treatment of ulcerative colitis based on network pharmacology and experimental validation

“…Frontiers in Pharmacology frontiersin.org (50 mg/kg) after intraperitoneal injection (Zhao et al, 2022). Five milliliters of abdominal aortic blood were collected and placed into the test tube, centrifuged at 3,000 r/min at 4 °C for 15 min, and the separated supernatant was placed into the refrigerator at −80 °C for testing.…”